Identification and mapping of human saphenous vein medial smooth muscle proteins by two-dimensional polyacrylamide gel electrophoresis

Changing smooth muscle phenotype and abnormal cell proliferation are important features of vascular pathology, including the failure of saphenous vein bypass grafts. We have characterised and mapped protein expression in human saphenous vein medial smooth muscle, using two-dimensional (2-D) polyacrylamide gel electrophoresis. The 2-D system comprised a nonlinear immobilised pH 3-10 gradient in the first dimension (separating proteins with isoelectric point values between pH 3-10), and 12%T total gel concentration sodium dodecyl sulphate polyacrylamide gel electrophoresis in the second dimension (separating proteins in the range 14,000-200,000 Daltons). Using a combination of peptide mass fingerprinting by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry and partial amino acid sequencing by nanospray tandem mass spectrometry, a subset of 149 protein spots was analysed, with 129 protein spots being identified and mapped. The data presented here are an important addition to the limited knowledge of venous medial smooth muscle protein expression in vivo. Our protein map will facilitate the identification of proteins differentially expressed in human saphenous vein bypass grafts. In turn, this may lead to the elucidation of molecular events involved in saphenous vein bypass graft failure. The map should also provide a basis for comparative studies of protein expression in vascular smooth muscle of varying origins.     

Species relationships and population structure of Littorina saxatilis Olivi and L. tenebrosa Montagu in Ireland using single-strand conformational polymorphisms (SSCPs) of cytochrome b fragments

Littorina saxatilis is a ubiquitous snail of intertidal habitats in the North Atlantic. Shell type in littorinids is extremely polymorphic and defined by habitat. Taxonomy based upon shell type has been revised in the light of anatomic and genetic information, but uncertainties remain. In this study, the population structure of L. saxatilis and L. tenebrosa was studied at 11 sites in Ireland using single-strand conformational polymorphisms of a 375-bp portion of the cytochrome b gene, and the status of L. tenebrosa, the small, fragile-shelled, brackish water type, was considered. The genetic patterns among L. saxatilis and L. tenebrosa populations were examined over varying distances and L. tenebrosa was compared with adjacent L. saxatilis populations at four sites on the west coast of Ireland and one site on the east coast. Haplotype diversity was high with 32 haplotypes present among 995 individuals. Pairwise tests suggest gene flow over small scales among and between habitat types and may reflect the stochastic legacy of postglacial recolonization over larger scales. In AMOVA tests, geography explained nearly twice as much of the variance (30%) as habitat type (18%), indicating that gene flow is more restricted by distance than by habitat type, and supporting the status of L. tenebrosa as an ecotype of L. saxatilis rather than a separate species.     

Risks of Hemolysis in Glucose-6-Phosphate Dehydrogenase Deficient Infants Exposed to Chlorproguanil-Dapsone,Mefloquine and Sulfadoxine-Pyrimethamine as Part of Intermittent Presumptive Treatment of Malaria in Infants

Background

Chlorproguanil-dapsone (CD) has been linked to hemolysis in symptomatic glucose-6-phosphate dehydrogenase deficient (G6PDd) children. Few studies have explored the effects of G6PD status on hemolysis in children treated with Intermittent Preventive Treatment in infants (IPTi) antimalarial regimens. We sought to examine the joint effects of G6PD status and IPTi antimalarial treatment on incidence of hemolysis in asymptomatic children treated with CD, sulfadoxine-pyrimethamine (SP), and mefloquine (MQ).

Methods

A secondary analysis of data from a double-blind, placebo-controlled trial of IPTi was conducted. Hemoglobin (Hb) measurements were made at IPTi doses, regular follow-up and emergency visits. G6PD genotype was determined at 9 months looking for SNPs for the A- genotype at coding position 202. Multivariable linear and logistic regression models were used to examine hemolysis among children with valid G6PD genotyping results. Hemolysis was defined as the absolute change in Hb or as any post-dose Hb <8 g/dL. These outcomes were assessed using either a single follow-up Hb on day 7 after an IPTi dose or Hb obtained 1 to 14 or 28 days after each IPTi dose.

Findings

Relative to placebo, CD reduced Hb by approximately 0.5 g/dL at day 7 and within 14 days of an IPTi dose, and by 0.2 g/dL within 28 days. Adjusted declines in the CD group were larger than in the MQ and SP groups. At day 7, homo-/hemizygous genotype was associated with higher odds of Hb <8 g/dL (adjusted odds ratio = 6.7, 95% CI 1.7 to 27.0) and greater absolute reductions in Hb (-0.6 g/dL, 95% CI -1.1 to 0.003). There was no evidence to suggest increased reductions in Hb among homo-/hemizygous children treated with CD compared to placebo, SP or MQ.

Conclusions

While treatment with CD demonstrated greater reductions in Hb at 7 and 14 days after an IPTi dose compared to both SP and MQ, there was no evidence that G6PD deficiency exacerbated the adverse effects of CD, despite evidence for higher hemolysis risk among G6PDd infants.     

An enzyme-linked immunosorbent assay (ELISA) for bovine LH capable of monitoring fluctuations in baseline concentrations

Using a Fab'-horseradish peroxidase conjugate, an ELISA for bovine LH was developed and validated. It had good specificity (less than or equal to 0.2% cross-reactivity with FSH or TSH), was not susceptible to interference from plasma components and exhibited very low non-specific binding. It was carried out on microtitre plates and had incubation times totalling 4 or 18 h, not including the initial antibody coating step. The longer incubations gave a lower detection limit (10 pg, 300 amol) and were used when fluctuations in baseline concentrations were being monitored. The values determined were independent of the plasma volume employed, standard added to plasma samples was accurately determined and the results obtained from the analysis of plasma samples correlated closely with those obtained by radioimmunoassay. The assay was applied to the analysis of plasma samples taken from heifers with normal ovarian activity or treated in a number of ways, and in every case the results obtained were similar to those determined by radioimmunoassay and reported in the literature.     

Scent marking and resource defence by male coypus (Myocastor coypus)

The alternative ideas that scent marking functions mainly in intrasexual competition or for attracting or stimulating mates were tested using data from a 16-year study of coypus, Myocastor coypus . Male coypus have a large anal gland, specialized for marking. Despite year-round breeding, the anal gland shows regular seasonal variation with marked enlargement in OctobeDecember. The analysis supported the hypothesis that this seasonal increase in glandular activity was linked to intrasexual competition: the size of the autumn peak was correlated with the increase in male numbers from the previous season and with the frequency of fighting scars. There were no relationships between gland size and any measures of the availability of mates. Increased scent marking may have been part of the response by resource-holding males to male recruits as these competed for existing territories. The timing of the autumn peak in marking and fighting may have been a response to the accumulation of non-resource-holding males over the summer (when recruitment exceeded mortality in the population) and to increasing accessibility within previously dense wetland habitats; together, these factors may cause a switch from scramble to despotic competition and an increase in scent marking to help keep the costs of resource defence within economic limits.     

A physical properties based approach for the exploration of a 4-hydroxybenzothiazolone series of β2-adrenoceptor agonists as inhaled long-acting bronchodilators

The chiral synthesis of a 4-hydroxybenzothiazolone based series of β₂-adrenoceptor agonists is described. Using this methodology a library of N-substituted analogues were prepared for the rapid identification of leads with the potential to be fast onset and long-acting inhaled bronchodilators with improved therapeutic margins. The design of the library to achieve the targeted profile was based upon lipophilicity and metabolism based hypotheses. This approach identified β-phenethyl, α-substituted cyclopentyl and monoterpene N-substituents to be of particular interest for further evaluation, as exemplified by structures 19, 29 and 33, respectively.     

Body odor similarity in noncohabiting twins

There is currently considerable interest in biometric approaches using human odor as a marker of disease or genetic individuality. Body odor is also thought to be used during mate choice to select genetically compatible mates. The idea that body odor reveals information about both genetic identity and genetic similarity is most readily tested by examining odor in twin pairs. However, although this idea can be traced back 130 years to Francis Galton in 1875, most studies using dogs fail to control for shared environmental effects associated with cohabitation. Here we show that odors of identical twins (but not dizygotic twins) can be matched by human sniffers at rates better than chance, even when the twins are living apart. In addition, matching frequencies for identical twin odors were not significantly different from those for duplicate odors from the same individual. These results indicate an important genetic influence on body odor and the potential for developing technologies for human odor printing in relation to underlying genotype.